High-throughput multi laser wave mixing detection methods and apparatus

ABSTRACT

This invention relates to methods and apparatus of a combination of multi-laser wave mixing technology with diagnostic flow technologies with embodiments describing capillary electrophoresis. The unique combination of these technologies along with minute detection levels not yet been seen in the field.

This application claims priority under 35 U.S.C. §119(e) to U.S.provisional patent application 61/523,538, filed Aug. 15, 2011.

This invention relates to methods and apparatus of a combination oflaser wave mixing technology with capillary electrophoresis diagnosticflow technologies. The combination of these technologies along withminute detection levels not yet been seen in the field.

BACKGROUND

Laser wave mixing has been described in many patents, journals andarticles. Having greatest relation to embodiments of the inventiondescribed herein are Tong has described degenerate four wave mixing andapparatus therein in U.S. Pat. Nos. 5,600,444 and 6,141,094 and PatentApplication 2006263777. These describe apparati and methods that intheir capacities are capable of analyzing small quantities of analytesdown to a detection level of attomoles. They utilize differentcomplements of analysis systems including HPLC and HCPE and a gas phaseatomizer type spectroscopy. Furthermore, the dissertation “ProteinAnalysis at the Single Cell Level by Nonlinear Laser Wave-MixingSpectroscopy for High Throughput Capillary Electrophoresis Applications”from Sadri's PhD dissertation N.C. State from 2008 relates similarapparati discussed in the Tong patents that reach the levels ofdetection of yoctomoles (10⁻²⁴). The named articles, dissertations andpatents are incorporated by reference in their entirety. Thesereferences give a background into the theories, adjustments andvariations upon the technology that are explanatory. Similarly,capillary electrophoresis (CE) has been explained and describe in manypatents and journal articles. A current review article gives a goodexample of the technology as used with peptides “Peptide Separation byCapillary Electrophoresis with Ultraviolet Detection: Some SimpleApproaches To Enhance Sensitivity and Resolution,” L. Noumie Suragau,Malaysian Journal of Analytical Sciences, 15:2 (2011)273-287. Thisreference gives a current view of CE technology with peptides as anexample analyte. Some advantages of CE are: employs capillary tubingwithin which the electrophoretic separation occurs; adaptable to moderndetector technology to give ease of use output; has great efficiencies;requires minute amounts of sample; easily automated for precisequantitative analysis and ease of use; consumes limited quantities ofreagents thus making it environmentally friendly; is applicable to awide selection of analytes.

As used in this specification and in the appended claims, the singularforms “a,” an” and “the” include plural references unless the contentclearly dictates otherwise.

The use of the word “preferably” in its various forms is explanatory forease of reading, and should not be used to read into the claims aslimiting or anything more.

In describing the invention and embodiments, the following terms will beemployed and are intended to be defined as indicated below. If any termsare not fully defined, then the normal usage as used in the art willfill any gaps in the understanding of the terminology.

Laser: is a device that creates a beam of light where all of the photonsare in a coherent state—usually with the same frequency and phase. Amongthe other effects, this means that the light from a laser is oftentightly focused and does not diverge much, resulting in the traditionallaser beam. In free space, the beams inside and outside the cavity areusually Gaussian distributed and are highly collimated with very smalldivergence. The distance over which the laser beam remains collimateddepends on the square of the beam diameter while divergence angle variesinversely with the beam diameter.

Collimating—is the process of making light rays parallel from a mixtureof diverging light rays or beams, and therefore will spread slowly as itpropagates. The word is related to “collinear” and implies light thatdoes not disperse with distance (ideally), or that will disperseminimally (in reality). A perfectly collimated beam with no divergencecannot be created due to diffraction, but light can be approximatelycollimated by a number of processes, for instance by means of acollimator or collimating lens.

Diagnostic flow technology: Is a solid state technology through a seriesof pumps or pump like mechanisms (such as electroosmotic flow,electrophoretic flow, capillary action, siphoning, pressure, implodinggas bubbles and the like) and apparati move analytes from a samplecollection area to an analysis area which comprise of multiple detectorstypes such as photodiode arrays (PDA), ultraviolet-visible (UV-VIS)spectrometers, charge coupled device (CCD) (such as a CCD-camera) massspectrometer (MS), Infrared spectrometers (such as Fourier TransformInfrared (FT-IR)], Nuclear Magnetic Resonance (NMR) detectors,Refractive Index spectrometers (RI), fluorescence detectors, radiationphotomultipliers, and the like. Flow can be achieved through liquids,fluids, gas or other means pumped or other means driven through a seriesof channels and mediums (such as tubing or silica gels) to move analytesfrom one point to another. Examples would comprise but not limited toLiquid Chromatography (LC) (which would further comprises variationssuch as micellar, ion exchange and the like), Reverse Phase HighPerformance Liquid Chromatography (RP-HPLC), Gas Chromatography (GC),High Performance Capillary Electrophoresis (HPCE), Capillary ZoneElectrophoresis (CZE), Supercritical Fluid Chromatography (SFC),Sub-critical Fluid Chromatography (SubFC), Inductively Coupled Plasma(ICP), and the like. Each technology is unique unto its own withpositives and negatives propagating from each in achieving the needs ofthe user. For example, capillary electrophoresis has environmentalpositives in utilizing very little hazardous materials but has negativeissues in what substances in what solvents are compatible.

Focal spot: an area or point onto which collimated light parallel to theaxis of a lens is focused. This spot of light can be expanded andcontracted in different shapes and geometries by some means such as acylindrical lens.

Absorptive interaction: interaction of analytes in a flow cell chamberor multi channel chamber when the two input beams are mixed and focusedin an absorbing medium. These beams form light induced gratings whenanalytes absorb the excitation light beam. The excited molecules in theform of interference patterns release their heat energy to surroundingsolvent or matrix molecules, creating dynamic thermal gratings, and as aresult, refractive index gratings. The incoming photons from the probebeam diffract off the gratings to generate the output signal beams.

Multichannel chamber: an enclosed space in which is configured to allowan absorptive interaction between multiple analytes and light beams.Multichannel flow cells and multiple capillary arrays can be situated ina multichannel chamber.

SUMMARY

The embodiments explained and described here utilize techniques toelucidate very small amounts of analyte with high sensitivity,selectivity, resolution and throughput.

The embodiments comprise of a diagnostic flow technology interconnected,configured with or linked to a multiple non linear optical wave mixingtechnique of a laser source of light absorptively interacting with ananalytes either in or passing through the multichannel chamber alsoknown as a laser sensing. Wherein, the interaction of the analyte andbeam of light are sensed by photodetectors to a very small molar amountthreshold.

The embodiments of the invention can be described by example. In asummary example, a device couples two a low watt quadruple Nd:YAG laserbeam in a unique ultraviolet (UV) wavelength of 266 nm utilizing a nonlinear wave mixing technique with a capillary electrophoresis diagnosticflow technology utilizing a capillary array. This example device can beused to elucidate concurrent multiple non-tagged or non-labeled nativeproteins that include in their sequence an amino acid picked from atleast three amino acid residues of tryptophan, tyrosine, andphenylalanine down to the levels of yoctomoles (10⁻²⁴) andsub-yoctomoles.

Embodiments reaching this yoctomole sensitivity allows for very smallinjected sample quantities. These levels would have many broad spectrumuses in pharmaceutical, environmental, forensic and anti-terrorismindustries. Analyzing such multiple small quantities can increaseefficiencies in time and cost in analysis procedures. The embodiments'configurations allow for short optical path lengths which can allow forcompact miniaturization of the equipment box. Embodiments of theinvention can achieve 100% optical collection efficiencies for signalsmeasured against a dark background.

Implementation of the embodiments comprise methods of analyzingsubstances through use of a diagnostic flow technology injecting a smallamount of analytes into a multichannel chamber, creating multiple beamsof light through the use of a non linear optical wave mixing technique,eliciting or generating a signal for each analyte, sensing the signalbeams, and manipulating and storing the data. Embodiments reaching thisyoctomole sensitivity allows for very small injected sample quantities.

An embodiment of the invention utilizes methods of analysis of thecombination of technologies. Included in these methods is creating asingle low watt laser beam also known as a light beam or light ray bysome laser sensing technology. From propagation the laser beam will beguided and manipulated through a series of devices, reflective surfacessuch as mirrors, beam traps, beam blockers, beam choppers, beamsplitters, focusing lenses, collimating lenses, and concave lenses withan interconnecting to electronic devices including, a computer to bothcontrol the front end processes of propagating and manipulating thelight source and running the diagnostic flow technology to the back endprocess of receiving the data and processing it into useable output.Electronics included are photodetectors such as a photodiode detector,an N-type Metal Oxide Semiconductor (NMOS) linked to a photodiode array(PDA) image detector or sensor, to receive the signal light input whichcould include an amplification of the signal with a photo multipliertube, a lock in amplifier to filter out extraneous frequencies, a beamchopper controller which controls or segregates the frequency in whichthe output beam is settled.

As the beam is split with a 70:30 ratio into two beams, the beams arethen focused onto a target area of the capillary window in the multichannel chamber where then a cylindrical lens expands the light wave tocover all the capillaries in the capillary arrays. This multi channelchamber is the interaction and interconnection of the diagnostic flowtechnology with the laser wave mixing. In one embodiment the diagnosticflow technology is an analytical CE device. This device has a source ofhigh voltage with microbore multi capillaries interconnected to anelectrophoretic buffer solutions with platinum cathode and anelectrophoretic buffer solutions with platinum anode. Other embodimentsmay have a mass spectrum device connected to the fluidic capillary. Thesample interacts with convergent or divergent light beams moving throughthe target area aperture in the capillary array. After penetrating thecapillary array the diffracted signal beams are collimated into acoherent light beam. Other light diffractions and rays are captured in abeam trap. This beam is directed to a beam splitter with the beams sentto photo detector in some embodiments could be photodiode detector andNMOS PDA. The beams are detected and the signal is translated andprocessed through computer applications to useable data.

BRIEF DESCRIPTION OF DRAWINGS

The patent or application file contains at least one drawing executed incolor. Copies of this patent or patent application publication withcolor drawing will be provided by the Office upon request and payment ofthe necessary fee.

The objects, advantages, and features of the invention will become moreapparent from the following detailed description, when read inconjunction with the accompanying drawing, in which:

FIG. 1. is a schematic of an example embodiment of the invention showingthe guided pathway of the dual laser light beams in black and red fordistinction and clarity with the light beam interconnected to adiagnostic flow device capillary electrophoresis.

FIG. 1 a. is a schematic blow up of the multichannel stage showing aside view of square capillary array. Note that the right side of thefigure shows an expansion of the light beam entering the array and theleft side shows collinear signal beams leaving the array (does notrepresent true nature of light beams)

FIG. 1 b. shows a facial flat planar view of the front of the capillaryarray window and the capillaries jutting out transverse. Note the shapesused for beams are not necessarily representative of the correct angleof attack on the capillary window.

DETAILED DESCRIPTION

Referring to the embodiments in FIG. 1, a schematic view showing anembodiment of the invention utilizing a capillary array connected to aCE diagnostic flow technology. Two or more laser light sources arecontemplated by the embodiments of the invention. Each of the two laserlight sources 100 a and 100 b emits and presents coherent beams 110 aand 110 b to a beam splitter 120. Many sources of laser light arecontemplated but lower wattage lasers give advantages to cheaper priceand less robust materials in the beam manipulative devices. Preferredlaser is the quadrupled Nd:YAG laser emitting 266 nm radiation at a highpulse frequency. Embodiments contemplate different types of lasers.Depending on the techniques used in the cavity, such as Q-switching,mode locking or gainswitching, the laser output may be continuous wave(CW) or pulsed. When the waveform is pulsed, higher peak powers areachieved. Dye lasers and vibronic solid-state lasers can generate a widerange of wavelengths that are appropriate for generating extremely shortpulses of light (10⁻¹⁵ s). Other types of lasers contemplated are gassuch as Argon-ion, chemical, excimer, solid state, photonic crystal,semiconductor, free electron, bio, and exotic. A laser type forimplementation of the embodiments contemplated is a solid stateNeodynium: yttrium aluminum garnet (Nd:YAG) lasers tuned to 266 nmwavelength suitable for native protein absorption measurements. This UVlaser (Model, NU-10210-100, Teem Photonics, France) also offers lowpower consumption (5 mW) and a good beam quality. Embodiments of theinvention can use either higher power (>1 W) or lower power lasers (<1W). Lower power lasers allow for less damage to optical components, lesscost to acquire and to use. To prevent laser damage to opticalcomponents and depending on the wavelength ranges and power, there areseveral optical materials commonly used comprise of borosilicate crownglasses (BK7), UV grade fused Silica, CaF₂, MgF₂, crystal Quartz, Pyrexand Zerodur.

At beam split, the preferable split ratio of the laser beam is 70:30.Beams 130 a and 130 b travel to reflective surface or a mirror 150 whichbrings the beam to the beam chopper 170 controlled by chopper controller180 and lock-in amplifier 190 which among other things amplifies andmodulates the cycles of the light wave preferably to 200 Hz. Othercycles are contemplated as the utility demands. The modulated beam 200 aand 200 b travels to reflective surface or mirror 210 and redirects thebeams towards the focusing convex lens 220 preferably 10 cm. The beamsare focused onto the capillary window 410 as seen in FIG. 1 b of thetarget area on the capillary array on the multichannel chamber 250. Thetarget areas of each beam on the capillary window can be variable. Forexample purposes only, beam 200 a can be focused on a target area tothen be expanded to cover the bottom four capillaries of an eightcapillary array. Similarly beam 200 b can be focused on a target area tothen be expanded to cover the top 4 capillaries of an 8 capillary array.The two separate beams should have minimal overlap on the capillariesafter their expansion. After the target areas is focused upon, the beamsare expanded by cylindrical lens 230 to cover all the capillary tubes inthe array with little overlap. Similarly, the beams 140 a and 140 btravel to mirror 160 and redirects the beams towards similar focusingand expansion as the preferably lower ratio beam with the focusingconvex lens 220 and beam expansion cylindrical lens 230. The beams 140 aand 140 b should orient before the focusing lens roughly parallel withbeams 200 a and 200 b. The spatial configuration such as distance, sizeand shape of the lenses allows for the beam focusing and expansion whichallows for variable size focal spots and in variable areas on the X,Y,Zcoordinate plane of the capillary array window 410 as seen in FIG. 1 bsimilar in function to a flow cell in other applications on themultichannel chamber 250.

Dependent on the materials, type of laser, size of mirrors and lensesused embodiments of the invention may reach to yoctomoles level inanalysis of analytes with for merely an example of analyte of nativeprotein with an amino acid tyrosine in the sequence utilizing a laser atwavelength 266 nm.

Other analytes contemplated but not limited to are cells, biomoleculesand small molecules such as labeled or unlabelled tagged and un-taggedproteins, native proteins, peptides, peptidomimetics, polysaccharides,nucleic acids, amino acids, adjuvants, celluloses, biopolymericmolecules, lipids, cell parts, organic compounds, inorganic compounds,antibodies, DNA, RNA, variations on DNA and RNA, nucleotides, drug, drugcandidates, biopharmaceuticals, environmental chemicals, astralchemicals, geophysical chemicals, forensic chemicals, chiral,enantiomers, stereoisomers, optical isomers, solids, liquids and gases.At such low levels of concentration the real time analysis or efficientanalysis of metabolic chemicals are contemplated.

Contemplated wavelengths of the laser beam are from the belowultraviolet (UV) range through the visible light spectrum beyond theinfrared depending on the lasers capabilities and spectralcharacteristics of the analyte. For example, the UV spectrum for aminoacid residue tyrosyl, tryptophanyl, and phenylalanyl reaches a peak ofextinction coefficients between 245 nm and 280 nm. Native proteinsincluding L and D versions of the amino acids or residues would becontemplated examples of use of the UV spectrum detection. A laser beamtuned to a unique 266 nm wavelength would be efficient in absorbing ananalyte containing these residues Similarly in another example a proteinanalyzed with a laser beam tuned to 210 nm or 214 nm would efficientlyelucidate the peptide bond whose extinction coefficient reaches itsmaximum at 190 nm. Other embodiments contemplate UV wavelengths between10 nm and 400 nm, visible spectrum between 380 and 800 nm and infraredfrom 740 nm to 300000 nm. Embodiments contemplates individual UVwavelengths or spectrums of wavelengths ranging between 190 nm and 300nm with other individual UV wavelengths and ranges contemplated such as210 nm to 280 nm and an individual UV wavelength at 210 nm, 254 nm, 266nm, and 280 nm.

Now turning to FIG. 1A., the schematic cross-section view shows a blowup of the multichannel chamber 250 held on a rigid translational stagewith the view directly into the capillaries 238. The beams 200 a and 200b and 140 a and 140 b are focused then expanded and configured into abeam 240 a and 240 b for explanatory purposes of showing that theexpanded beams are covering in this case four upper capillaries by redbeam 240 a and the lower four capillaries by white beam 240 b as thedesired target area of the capillary window of the capillary arraysimilar to a sample cell window. The window should be stabilized andkept vibration free. The photons of the beams interact with the analytesamples flowing through a multichannel capillary window similar to aflow cell, in this embodiment, the multiple signal beams are merelyrepresented by the red beams 241 a and the white beams 241 b leave otherside of capillary window. The figure shows example beams as collinearbut it is not representative of true nature.

The expansion configured beams 240 a and 240 b is shown in FIG lb afront facial planar view of the capillary window 410 the window maybe ofvariable widths but preferably 0.5 cm of the capillary array 400. Thewindow has an array of eight capillaries 238 with the outer coveringremoved showing the naked capillary tubing. The four beams are shownentering the capillary window with the upper four capillaries with atransparent pinkish red hue expanded to cover all the capillaries in thewindow representing two beams 240 a′ and 240 a″ and the cleartransparent representing beams 240b′ an 240 b″. These are facialrepresentatives of the beams moving into the capillary to mix together.The angle of the beams entering is not representative.

Analytes are flowed and separated in the capillary array by means ofelectroosmotic and electrophorectic force by voltage from power supply220 applying a voltage across anode 220 a made from a proper materialsuch as platinum to cathode 220 b made from a proper material such asplatinum. Any variable amount of capillaries greater than 1 arecontemplated for embodiments of the invention. The capillaries may havevariable inner diameters (i.d.) and outer diameters (o.d.). The largernet o.d. of each capillary provides larger total capillary surface areaper array with larger distance between each capillary probe area. Thepreferred i.d. is 71 um. The capillaries can be made out of any chemicalcombination of materials to allow for flow of analyte into the samplestaging area and robust enough for any pressures the system would exerton them. The capillaries can be coated (such as polyimide) or uncoatedon the outer surface as the experiment demands. The coating should allowfor close proximity of the capillaries and allow for light penetration.The capillaries inner wall can be coated (such as polyacrylamide forvisible spectrum) or un-coated in the inner surfaces as the experimentdemands.

In embodiments utilizing CE, capillaries should be rinsed with waterbefore each run and filled up with a dynamic coating and sieving matrix.An example of a dynamic coating and sieving matrix is a solutioncomprising 50 mM TRIS borate, 2.5 mM EDTA, 0.5% methylcellulose (highviscosity), 5% Dextran and 0.1% SDS. Solutions should be transparent toapplied UV wavelengths.

The capillaries may have different shape geometries for example squareor round. The shape can allow among other things good bundling of thecapillaries, minimization of background optical noise, less opticalscattering and diffraction. The preferred shape is square configured toallow the least amount of gaps minimizing laser leakage between thecapillaries. The length of capillary can vary with an effective lengthbeing the side that brings the sample analyte to the capillary windowfor sensing and detection. A preferable effective length is 25 cm. Thenumber of capillaries can also be variable with the needs of theexperiment and limitations of the delivery system. The variable amountof the capillaries is greater than 1 such contemplated as 5, 6, 7, 8, 9,10, 11, 12 and greater than 12. The bundling configuration of thecapillaries can be in different 2 dimension or 3 dimension geometriesthat allow for the best penetration of light, less interference, opticalnoise, scattering and diffraction. For example, a flat stacked array ofcapillaries. Means of attaching of the capillaries would be uses ofglues, adhesives, or other such attachment means or through the packingconfiguration of the capillaries in a holder that needs no attachingmeans. The embodiments have the capability of variable focal point orspot of the beam interacting with the capillaries and can variably beadjusted to track the amount and configuration of the capillaries.

An example to summarize for use in an embodiment utilizing CE andanalyzing native unlabeled proteins is the capillaries would beun-coated on the outer surface, fused silica, utilizing a squaregeometry, an array amount of 10, configured in a stacked configurationand a coating transparent to UV on the inner surface with a 0.5 cmcapillary window.

Turning back to FIG. 1, the coherent remnant beams 245 a,b,c,d,e,f,g,hafter absorptive interaction in passing through the multichannel chamber250 the beams 245 a,b,c,d,e,f,g,h are separated into beams 245c,d,e,f,g,h into beam trap 270 and signal beam 245 a and 245 b to mirror280 which shifts the beams to a collimating lens 290 which among otherthings is used to prevent too much signal divergence and to minimizeoptical interference between capillaries. The beams 295 a and 295 b intoa beam splitter 300 in some ratio preferably 70:30. The beams 305 a and305 b are split to a photodiode detector 310 as a control and beams 305c and 305 d are split to a multi photospectrometer 320 preferably a NMOSPDA to be detected, stored and analyzed among other data manipulationsin the computer 330. It is contemplated analog to digital (A/D)converters would be used as needed by the application. The distance fromthe capillary window is important in bringing the beams to coherence andparallel without losing intensity.

While the invention has been described in terms of various preferredembodiments and specific examples, the invention should be understood asnot being limited by the foregoing detailed description, but as beingdefined by the appended claims and their equivalents.

What I claim is:
 1. A high throughput apparatus comprising multi laserwave-mixing sensing technology combined with a multichannel diagnosticflow technology.
 2. The apparatus of claim 1 wherein the diagnostic flowtechnology is a capillary array electrophoresis.
 3. The apparatus ofclaim 1, wherein the multichannel diagnostic flow technology comprises amulti array capillary electrophoresis and photodectors.
 4. The apparatusof claim 3, wherein the multi laser wave-mixing technology comprises: a.at least two UV laser sources, b. a guided pathway for the laser beams.5. The apparatus of claim 4, wherein the guided pathway for the laserbeams comprises of a series of devices to manipulate said laser beamsfurther comprising: a. a computer interconnected to electronic devices,b. a lock in amplifier, c. a beam chopper controller, d. a beam chopper,e. a beam splitter, f. a reflective mirror, g. a beam blocker, h. afocusing lens, i. a cylindrical lens, j. a beam trap, k. a secondaryreflective mirror, l. a collimating lens, m. a secondary beam blocker,n. a tertiary reflective mirror, o. a secondary beam splitter, p.photodetectors.
 6. The apparatus of claim 5, wherein the focusing lensis 10 cm diameter and the cylindrical lens is a UV fused silicacylindrical plano-concave lens.
 7. The apparatus of claim 3, wherein themulti array capillary electrophoresis comprises: a. a high voltagesource, b. an electrophoretic buffer, c. an anodic platinum electrode,d. a cathodic platinum electrode, e. microbore fused silica capillarytubing configured to connect the sample to the buffers and to acapillary array chamber, f. a multi sample injection port, g. acapillary array chamber.
 8. The apparatus of claim 7, wherein the fusedsilica is square shaped.
 9. The apparatus of claim 7, wherein the multiarray capillary chamber comprises of 10 square shaped fused silicacapillaries stripped of their outer coating 0.5 cm wide glued togetherin a flat plane creating a capillary window.
 10. The apparatus of claim9, wherein the effective length of the fused capillary is 25 cm.
 11. Theapparatus of claim 9, wherein the inner diameter of the fused capillaryis 71 um.
 12. The apparatus of claim 5, wherein the laser light beamswavelength is in the UV spectrum.
 13. The apparatus of claim 11, whereinthe laser light beam wavelength is 266 nm.
 14. The apparatus of claim 5,wherein the photodetectors comprise a NMOS photodiode array and aphotodiode detector.
 15. The apparatus of claim 5, wherein thecollimating lens is placed after the flow cell and before the secondarybeam blocker.
 16. A high throughput method comprising of steps: a.creating at least two low watt laser beams b. manipulating the laserbeams towards a multi array capillary chamber, c. charging cathodic andanodic buffer solutions, d. sampling multiple minute scale analytes, e.electrophorecticly flowing an analyte into the capillary aperture, f.focusing beams on small area target of capillary array window, g.expanding beams with minimal overlap until full coverage of allcapillaries in window, h. collecting divergent beams after penetrationinto flow cell i. manipulating signal laser beams towards a photodiodedetector and photodiode array, j. processing signals into useable data.17. The method of claim 16, wherein the laser beams are created by a lowwatt frequency quadrupled Nd:YAG laser.
 18. The method of claim 17wherein the minute scale analytes as passed through the target capillarywindow are analyzed at yoctomole concentration.
 19. The method of claim16 wherein the analytes are a native proteins further including at leastone amino acid chosen from the group consisting of L-phenylalanine,Ltryptophan, L-tyrosine, D-phenylalanine, D-tryptophan, and D-tyrosine.20. A high throughput apparatus comprising: a. a computer interconnectedto electronic devices, b. a 266 nm wavelength Nd:YAG laser, c. a second266 nm wavelength Nd:YAG laser, d. a guided pathway for light beamsfurther comprising, a lock in amplifier, a beam chopper controller, abeam chopper, a beam splitter set to ratio 70:30, a reflective mirror, abeam blocker, a 10 cm focusing lens, a UV fused silica cylindricalplano-concave lens, a beam trap, a secondary reflective mirror, acollimating lens, a secondary beam blocker, a tertiary reflectivemirror, a photodiode detector and a photodiode array, e. a CEinterconnected to the apparatus through a capillary array sample targetarea further comprising a high voltage source, an electrophoreticbuffer, f. platinum electrodes as a cathode and anode, g. microborefused silica capillary tubing configured to connect the sample to thebuffers and to the capillary array chamber, h. a multi sample injectionport, i. a multi array capillary chamber further comprising of aneffective length of 25 cm of 10 square shaped fused silica capillarieswith an inner diameter of 71 um stripped of their outer coating 0.5 cmwide glued together in a flat plane creating a capillary window.